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Identification, characterization, and expression analysis of a serotonin receptor involved in the reproductive process of the Pacific abalone, Haliotis discus hannai

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Abstract

Serotonin receptor (5-HT) is a biogenic amine acting as a neurotransmitter and neuromodulator that mediates various aspects of reproduction and gametogenesis. The full-length nucleotide sequence of Haliotis discus hannai encodes a protein of 417 amino acids with a predicted molecular mass of 46.54 kDa and isoelectric point of 8.94. The structural profile of 5-HTHdh displayed key features of G protein-coupled receptors, including seven hydrophobic transmembrane domains, putative N-linked glycosylation sites, and several phosphorylation consensus motifs. It shares the highest homology of its amino acid sequence with the 5-HT receptor from Haliotis asinina, and to lesser extent of human 5-HT receptor. The cloned sequence possesses two cysteine residues (Cys-115 and Cys-193), which are likely to form a disulfide bond. Phylogenetic comparison with other known 5-HT receptor genes revealed that the 5-HTHdh is most closely related to the 5-HTHa receptor. The three-dimensional structure of the 5-HTHdh showed multiple alpha helices which is separated by a helix-loop-helix (HLH) structure. Quantitative PCR demonstrated that the receptor mRNA was predominantly expressed in the pleuropedal ganglion. Significant differences in the transcriptional activity of the 5-HTHdh gene were observed in the ovary at the ripening stage. An exclusive expression was detected in pleuropedal ganglion, testis, and ovary at higher effective accumulative temperature (1000 °C). In situ hybridization showed that the 5-HTHdh expressing neurosecretory cells were distributed in the cortex of the pleuropedal ganglion. Our results suggest that 5-HTHdh synthesized in the neural ganglia may be involved in oocyte maturation and spawning of H. discus hannai.

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Acknowledgements

This research was a part of the project titled ‘Development of technology for abalone aquaculture using sperm cryopreservation (Grant No. 2018-2129)’ funded by the Ministry of Oceans and Fisheries, Korea.

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  1. Department of Fisheries Science, College of Fisheries and Ocean Sciences, Chonnam National University, 50 Daehak-ro, Yeosu, Jeonnam, 59626, Republic of Korea

    Md. Rajib Sharker, Zahid Parvez Sukhan, Soo Cheol Kim, Won Kyo Lee & Kang Hee Kho

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Kang Hee Kho supervised and wrote the manuscript, Md. Rajib Sharker executed the experiments and wrote the manuscript. Zahid Parvez Sukhan and Soo Cheol Kim assisted to perform the experiments and analyze the data. Won Kyo Lee did critical discussion with the manuscript.

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Supplementary material 1 (DOCX 57 kb) Supplementary file 1 Semi-quantitative RT (reverse transcription)-PCR expression of serotonin receptor in Pacific abalone tissues. RPL-5 was used as an internal control for normalization of mRNA expression in tissues.

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Sharker, M.R., Sukhan, Z.P., Kim, S.C. et al. Identification, characterization, and expression analysis of a serotonin receptor involved in the reproductive process of the Pacific abalone, Haliotis discus hannai. Mol Biol Rep 47, 555–567 (2020). https://doi.org/10.1007/s11033-019-05162-2

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